|Prenylation||Sumoylation||Tyrosine Sulfation||Tyrosine Nitration|
1. If the antigen is provided by the customer: modified peptide (<15 aa >90% purity, 24 mg) and unmodified control (<15 aa >90% purity, 11 mg) are needed.
Accord to our years of experience, the success rate of the modified antibody preparation is lower than the normal one. To ensure the probability of obtaining a useful antibody after immunization, we recommend to immunize 3-4 rabbits.
Modified synthetic peptide usually has a chain of 14 amino acids. Approximately 10 mg are used in modified epitope affinity purification, 10 mg are used in antibody production, and 4 mg are used in QC.
Approximately 10 mg of non-modified peptide (about 14 amino acids) are used in affinity purification.
To obtain a more specific modified antibody, sometimes we used shortened modified peptides to purify.
Proper design of a suitable antigen is the most critical step in getting a useful antibody.
Through professional analysis, we choose the best region to synthesize peptide coupling KLH as the antigen to immune animals.
Each synthesized peptide goes through HPLC and MS tests to ultimately obtain a useful antibody.
ABclonal offers DNA methylation, RNA methylation, pan-acetylation, non-histone modification and other custom specific modified antibodies that are developed to meet your research needs.
Use Nipponbare callus (same quantity, about 3 g) as sample material of ChIP experiment, 02g and 03g as target gene. Conduct IP experiment use anti-H3K27me3 (ABclonal) and anti-H3K27me3 (Company M).
The efficiency of ABclonal anti-H3K27me3 immunoprecipitation is 10 times that of Company M anti-H3K27me3.
Obtaining a useful site-specific antibody is not easy. Most manufacturers cannot achieve this because of antibody validation. In addition to manufacturing useful site-specific antibodies, we dedicate ourselves to be the best validation platform in the world. ABclonal has built advanced testing platforms for Western Blot, immunohistochemistry, and immunofluorescence-based assays, supplemented by immunoprecipitation, RIP, ChIP, ChIP-seq, and shRNA knockout validation. We take every measure to provide the most accurate and reliable data for each of our customers.
ABclonal's standard protocol for site-specific antibody is optimized to produce antiserum in the shortest possible time. It is the most affordable option and a popular choice among researchers.
The protocol uses two New Zealand White rabbits that are Specific-Pathogen-Free. Injections are subcutaneous (SQ) as emulsions in Complete Freund's Adjuvant (CFA) or Incomplete Freund's Adjuvant (IFA). We guarantee that the same antigen will be injected into both animals.
|Control Serum Collection||Day 0||Pre-immune bleed (5 mL per rabbit)|
|Primary Injection||Day 1||Immunize with 0.50 mg of antigen in CFA, 10 SQ sites|
|1st Booster||Day 14||Boost with 0.25 mg of antigen in IFA, 4 SQ sites|
|2nd Booster||Day 28||Boost with 0.25 mg of antigen in IFA, 4 SQ sites|
|Serum Collection||Day 35||Bleed (-25 mL per rabbit)|
|3rd Booster||Day 42||Boost with 0.25 mg of antigen in IFA, 4 SQ sites|
|Serum Collection||Day 56, 58||Two bleeds (total of -50 mL per rabbit)|
|ELISA Test||Day 60|| ELISA titration (results available online)
Verify disposition of rabbits
Decide to continue or terminate
|Purification||Optional||6-10 mg of purified antibodies|